Abstract
PurposeThe aim of this study was to analyze systematically the influence of the relative centrifugation force (RCF) on leukocytes, platelets and growth factor release within fluid platelet-rich fibrin matrices (PRF).Materials and methodsSystematically using peripheral blood from six healthy volunteers, the RCF was reduced four times for each of the three experimental protocols (I–III) within the spectrum (710–44 g), while maintaining a constant centrifugation time. Flow cytometry was applied to determine the platelets and leukocyte number. The growth factor concentration was quantified 1 and 24 h after clotting using ELISA.ResultsReducing RCF in accordance with protocol-II (177 g) led to a significantly higher platelets and leukocytes numbers compared to protocol-I (710 g). Protocol-III (44 g) showed a highly significant increase of leukocytes and platelets number in comparison to -I and -II. The growth factors’ concentration of VEGF and TGF-β1 was significantly higher in protocol-II compared to -I, whereas protocol-III exhibited significantly higher growth factor concentration compared to protocols-I and -II. These findings were observed among 1 and 24 h after clotting, as well as the accumulated growth factor concentration over 24 h.DiscussionBased on the results, it has been demonstrated that it is possible to enrich PRF-based fluid matrices with leukocytes, platelets and growth factors by means of a single alteration of the centrifugation settings within the clinical routine.ConclusionsWe postulate that the so-called low speed centrifugation concept (LSCC) selectively enriches leukocytes, platelets and growth factors within fluid PRF-based matrices. Further studies are needed to evaluate the effect of cell and growth factor enrichment on wound healing and tissue regeneration while comparing blood concentrates gained by high and low RCF.
Highlights
In recent years, several concepts have been introduced for clinically relevant tissue engineering by means of minimally invasive approaches
The third protocol-III (44 g) with four times less relative centrifugation force (RCF) than protocol-II and 16 times less RCF than protocol-I revealed the highest number of leukocytes, which was statistically highly significant compared to protocol-I (P < 0.0001) and protocol-II (P < 0.0001) (Fig. 1a)
The results showed that decreasing RCF up to 16 times less than the first described PRF led to a significant increase in leukocyte and platelets numbers within the generated PRF-matrices
Summary
Several concepts have been introduced for clinically relevant tissue engineering by means of minimally invasive approaches. Well-accepted models include pure biomaterial application [1,2,3,4] and the combination of biomaterials with autologous bone marrow aspirates [5, 6] or recombinant bone morphogenetic proteins (BMPs) [7, 8]. Less invasive than autologous bone transplantation, the previously mentioned minimally invasive bone marrow aspirate methodologies are still reserved for only experienced surgeons, as these methods can be associated with complications such as pain,. Damage to the adjacent organs during bone marrow retrieval. The optimal growth factor concentration needs to be determined for the use of BMPs
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